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高等学校化学研究 ›› 2014, Vol. 30 ›› Issue (2): 272-278.doi: 10.1007/s40242-014-3311-z

• Articles • 上一篇    下一篇

Proteomics Dissection of Cold Responsive Proteins Based on PEG Fractionation in Arabidopsis

LI Shanyu2, LIU Xiangguo3, WANG Shang1, HAO Dongyun3, XI Jinghui1   

  1. 1. College of Plant Science, Jilin University, Changchun 130062, P. R. China;
    2. The First Hospital of Jilin University, Changchun 130021, P. R. China;
    3. Biotechnology Research Centre, Jilin Academy of Agricultural Sciences, Changchun 130124, P. R. China
  • 收稿日期:2013-07-18 修回日期:2013-11-15 出版日期:2014-04-01 发布日期:2013-11-25
  • 通讯作者: XI Jinghui E-mail:jhxi1965@jlu.edu.cn
  • 基金资助:

    Supported by the National Natural Science Foundation of China(No.30470159), the Public Benefit Research Foundation of Ministry of Agriculture of China(No.201003025) and the World Bank Loan Project from Department of Finance of Jilin Province, China(No.2011-Z19).

Proteomics Dissection of Cold Responsive Proteins Based on PEG Fractionation in Arabidopsis

LI Shanyu2, LIU Xiangguo3, WANG Shang1, HAO Dongyun3, XI Jinghui1   

  1. 1. College of Plant Science, Jilin University, Changchun 130062, P. R. China;
    2. The First Hospital of Jilin University, Changchun 130021, P. R. China;
    3. Biotechnology Research Centre, Jilin Academy of Agricultural Sciences, Changchun 130124, P. R. China
  • Received:2013-07-18 Revised:2013-11-15 Online:2014-04-01 Published:2013-11-25
  • Contact: XI Jinghui E-mail:jhxi1965@jlu.edu.cn
  • Supported by:

    Supported by the National Natural Science Foundation of China(No.30470159), the Public Benefit Research Foundation of Ministry of Agriculture of China(No.201003025) and the World Bank Loan Project from Department of Finance of Jilin Province, China(No.2011-Z19).

摘要:

Proteome profiling was performed on Arabidopsis plant exposed to cold stress at 4 ℃ for 24 h in an attempt to explore the mechanisms of plant climate adaptation. The polyethylene glycol(PEG) fractionation protocol developed in this lab was used to identify as many differentially expressed low-abundance proteins as possible. In comparison with those of the biological controls, 67 protein spots with at least two-fold difference in expression were identified for the plant exposed to cold temperatures; and from these spots, 50 proteins were successfully identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS). Bioinformatics studies on these proteins show that 57.8% of these proteins were localized in the chloroplast. Of these proteins, 8 ones have functions in photosynthesis, including glycine hydroxymethyltransferase, Rubisco large subunit, Rubisco activase, PSBO2, fructose-1,6-bisphosphate aldolase, NADP-dependent malate dehydrogenase, sedoheptulose bisphosphatase and photosystem Ⅱ reaction center PsbP family protein, suggesting that photosynthesis is greatly affected by cold stress. The identified proteins were validated by quantitative real-time polymerase chain reaction(qPCR). Taken together, our results suggest that the chloroplast might also act as a cold stress sensor and that photosynthesis-related proteins may play important roles in cold acclimation for Arabidopsis.

关键词: Arabidopsis thaliana, Cold stress, Proteome, Protein fractionation

Abstract:

Proteome profiling was performed on Arabidopsis plant exposed to cold stress at 4 ℃ for 24 h in an attempt to explore the mechanisms of plant climate adaptation. The polyethylene glycol(PEG) fractionation protocol developed in this lab was used to identify as many differentially expressed low-abundance proteins as possible. In comparison with those of the biological controls, 67 protein spots with at least two-fold difference in expression were identified for the plant exposed to cold temperatures; and from these spots, 50 proteins were successfully identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS). Bioinformatics studies on these proteins show that 57.8% of these proteins were localized in the chloroplast. Of these proteins, 8 ones have functions in photosynthesis, including glycine hydroxymethyltransferase, Rubisco large subunit, Rubisco activase, PSBO2, fructose-1,6-bisphosphate aldolase, NADP-dependent malate dehydrogenase, sedoheptulose bisphosphatase and photosystem Ⅱ reaction center PsbP family protein, suggesting that photosynthesis is greatly affected by cold stress. The identified proteins were validated by quantitative real-time polymerase chain reaction(qPCR). Taken together, our results suggest that the chloroplast might also act as a cold stress sensor and that photosynthesis-related proteins may play important roles in cold acclimation for Arabidopsis.

Key words: Arabidopsis thaliana, Cold stress, Proteome, Protein fractionation